Addition of collagen type I in agarose created a dose-dependent effect on matrix production in engineered cartilage.

Extracellular-matrix composition impacts mechanical performance in native and engineered tissues. Previous studies showed collagen type I-agarose blends increased cell-matrix interactions and extracellular matrix production. However, long-term impacts on protein production and mechanical properties of engineered cartilage are unknown. Our objective was to characterize the effect of collagen type I on the matrix production of chondrocytes embedded in agarose hydrogels. We hypothesized that the addition of collagen would improve long-term mechanical properties and matrix production (e.g. collagen and glycosaminoglycans) through increased bioactivity. Agarose hydrogels (2% w/v) were mixed with varying concentrations of collagen type I (0, 2 and 5 mg/ml). Juvenile bovine chondrocytes were added to the hydrogels to assess matrix production over 4 weeks through biochemical assays, and mechanical properties were assessed through unconfined compression. We observed a dose-dependent effect on cell bioactivity, where 2 mg/ml of collagen improved bioactivity, but 5 mg/ml had a negative impact on bioactivity. This resulted in a higher modulus for scaffolds supplemented with lower collagen concentration as compared to the higher collagen concentration, but not when compared to the control. In conclusion, the addition of collagen to agarose constructs provided a dose-dependent impact on improving glycosaminoglycan production but did not improve collagen production or compressive mechanics.

Agarose-Based Hydrogels as Suitable Bioprinting Materials for Tissue Engineering.

Hydrogels are useful materials as scaffolds for tissue engineering applications. Using hydrogels with additive manufacturing techniques has typically required the addition of techniques such as cross-linking or printing in sacrificial materials that negatively impact tissue growth to remedy inconsistencies in print fidelity. Thus, there is a need for bioinks that can directly print cell-laden constructs. In this study, agarose-based hydrogels commonly used for cartilage tissue engineering were compared to Pluronic, a hydrogel with established printing capabilities. Moreover, new material mixtures were developed for bioprinting by combining alginate and agarose. We compared mechanical and rheological properties, including yield stress, storage modulus, and shear thinning, as well as construct shape fidelity to assess their potential as a bioink for cell-based tissue engineering. The rheological properties and printability of agarose-alginate gels were statistically similar to those of Pluronic for all tests (p > 0.05). Alginate-agarose composites prepared with 5% w/v (3:2 agarose to alginate ratio) demonstrated excellent cell viability over a 28-day culture period (>∼70% cell survival at day 28) as well matrix production over the same period. Therefore, agarose-alginate mixtures showed the greatest potential as an effective bioink for additive manufacturing of biological materials for cartilage tissue engineering.